The rising concern of multidrug-resistant pathogens and the unavailability of new antibiotics in the discovery pipeline has generated the need for alternative approaches for their control. Essential oils (EOs) and essential oil's active components (EoCs) have been reported for their antimicrobial effect since long. In this study, we evaluated 11 EOs and 4 EoCs for their antimicrobial and antibiofilm activity. Among EOs, oregano (zone of inhibition [ZOI] = 24.62 ± 8.10 mm), cinnamon (ZOI = 18.38 ± 5.88 mm), cassia (ZOI = 20.23 ± 4.61 mm), and thyme (ZOI = 13.77 ± 5.67 mm), and among EoCs, cinnamaldehyde (ZOI = 24.69 ± 5.83 mm), carvacrol (ZOI = 24.38 ± 7.76 mm), and thymol (ZOI = 22.92 ± 7.43 mm) demonstrated the higher antimicrobial activity against the targeted pathogens. Only cinnamon (ZOI = 16.62 ± 2.58 mm) and cassia (ZOI = 15.24 ± 1.90 mm) were found to be effective as antimicrobial against Pseudomonas aeruginosa strains. Cinnamaldehyde (ZOI = 9.02 ± 2.68 mm) demonstrated weak antimicrobial activity. Minimum inhibitory concentration (MIC), minimum bactericidal concentration (MBC), and antibiofilm assays were performed in triplicate. Statistical analysis using the Kruskal-Wallis test revealed a significant overall difference between treatments (p < 0.05) against both P. aeruginosa and Staphylococcus aureus. Specifically, Dunn's multiple comparisons test showed significant differences (p < 0.05) in MIC values between cinnamaldehyde and eugenol for P. aeruginosa with no significant difference in their MBC. For S. aureus, significant variations (p < 0.05) in MIC and MBC were observed between carvacrol and thymol, and between thyme and other oils like tea tree, cumin, and lemongrass. Antibiofilm effect of EOs and EoCs against P. aeruginosa and S. aureus was found at concentration of 0.031% for cinnamon EO and cinnamaldehyde and at 0.015 - 0.007% for carvacrol. Gas chromatography-mass spectrometry analysis of EOs and EoCs was performed to determine their bioactive compounds. The study highlights that EOs and EoCs can be used as alternative antimicrobials against food spoilage bacteria and drug-resistant pathogens.
Foodborne diseases continue to be major public health challenges around the world, with contaminated food linked to an estimated 420,000 deaths annually, many of which are children under five. Traditional testing methods for foodborne pathogens (i.e., microbial culturing and biochemical assays), while generally reliable, are slow and labor-intensive and significantly limit their ability to protect the public from exposure to foodborne pathogens. Reflecting on these limitations, a growing number of methods using rapid detection approaches, which are more efficient for detecting foodborne pathogens, have been developed, including nucleic acid-based methods, immunological methods, and biosensor-based methods. In particular, multiplex polymerase chain reaction (mPCR) offers a good method for the rapid detection of multiple pathogens in food samples. This review highlighted both the progress and prospects of multiplex-allele-specific PCR (MAS-PCR) technology and its use to detect and genotype foodborne pathogens in food samples. Also, the MAS-PCR used with probe-based methods has the possibility of being more specific and sensitive than the single PCR assay alone. The subsequent new approaches are designed to minimize the time it takes to identify pathogens to improve food safety and avoid disease outbreaks. Ultimately, advanced technologies would demonstrate increased speed and accuracy of pathogen detection, creating a greater contribution towards global food safety and public health.
Foodborne pathogens represent a significant public health burden. Quantifying the relative importance of various potential sources of foodborne infection is challenging due to data scarcity and uncertainty in empirical studies. Structured expert judgment (SEJ) provides a valuable methodological alternative to gain insights into source attribution of foodborne pathogens. We conducted a SEJ study to attribute human cases of 26 foodborne pathogens in the Netherlands to seven major transmission pathways, 20 food groups, and two animal groups, in a typical post-COVID19 year, using Cooke's classical model. The elicitation process involved snowball recruitment, expertise self-assessment, and a workshop where experts answered calibration questions to capture their uncertainty as input for the model. Subsequently, experts completed the target questions to obtain attributable proportions at the 'kitchen door' level. Results indicated that transmission was predominantly (>50%) foodborne for Staphylococcus aureus, Listeria monocytogenes, Yersinia spp., Bacillus cereus, Clostridium perfringens, certain non-typhoidal Salmonella serotypes, Campylobacter spp., hepatitis E virus and Toxoplasma gondii, whereas person-to-person transmission was the primary pathway for astrovirus, rotavirus, norovirus, and sapovirus. Brucella spp. and typhoidal Salmonella were attributed primarily (>85%) to international travel. All other pathogens showed attributions of <50% to any individual pathway. Substantial differences were observed when dividing foodborne transmission into food groups. Key contributors included food handlers and vermin, various meats (e.g., pork, beef, chicken), and shellfish. These SEJ-derived estimates complement existing data by providing pathogen-specific insights in the Dutch context.
Foodborne diseases pose a significant public health and economic burden worldwide. This study systematically analyzed a decade (2014-2023) of national surveillance data from mainland China, encompassing 50,434 outbreaks, 311,672 illnesses, and 1330 fatalities. The results revealed pronounced geographical disparities, with five provinces (Shandong, Yunnan, Hunan, Guizhou, and Sichuan) collectively accounting for 59.27% of outbreaks. A distinct seasonal peak was observed from June to August, correlated with high temperature and humidity. Poisonous mushrooms were the predominant causative agent, responsible for 50.65% of outbreaks with identified etiology and the highest case fatality. Microbial pathogens, notably Vibrio parahaemolyticus and Salmonella spp., were also major contributors. Most outbreaks and fatalities occurred in household settings, with a notable surge during the COVID-19 pandemic. Our findings underscore the critical need for targeted interventions, enhanced public education on toxic mushrooms, and strengthened surveillance systems to reduce the burden of foodborne diseases in China.
This study aimed to perform laboratory testing and traceability analysis of suspicious samples from a foodborne disease outbreak caused by Clostridium perfringens (C. perfringens) in a hospital in Ma'anshan City, Anhui Province, and to evaluate the application of whole-genome sequencing technology in the tracing of foodborne disease outbreaks. On-site epidemiological investigations were conducted, and suspicious biological, food, and environmental samples were collected. Patient biological samples underwent multi-pathogen screening. Following the epidemiological analysis, isolation, culture, and identification of suspected pathogens were carried out. Toxin gene detection and whole-genome sequencing were then performed on the isolated C. perfringens strains. A total of 131 samples were collected during the outbreak. Among these, 20 anal swabs from patients with diarrhoea were tested for the nucleic acids of foodborne pathogens and five types of Diarrheagenic Escherichia coli. All samples tested positive for C. perfringens nucleic acid, while six samples also tested positive for enteropathogenic E. coli. Isolation and culture revealed that C. perfringens was detected in 41 samples, with a detection rate of 31.3% (41/131). Nucleic acid detection of six toxin genes (plc, cpb, cpe, Ia, etx, and netB) was performed on the 41 C. perfringens strains, with 29 strains testing positive for cpe. Whole-genome sequencing of the cpe-positive strains revealed identical ST sequence types and a maximum of 25 SNP differences, indicating a highly clonal group. Phylogenetic analysis of the core genome demonstrated homology among C. perfringens strains from patient anal swabs, canteen employees's anal swabs, and food samples. The outbreak was likely caused by contamination of Chinese food by C. perfringens from infected canteen employees. Whole-genome sequencing was instrumental in accurately tracing the source of the outbreak.
Foodborne and waterborne gastrointestinal disease outbreaks are frequently reported from the northeastern region of India. In the present study, an outbreak of acute gastroenteritis in Keifang village of Mizoram, India, was investigated following official notification through the state Integrated Disease Surveillance Program (IDSP) unit. The investigation involved descriptive epidemiology and exposure assessment for 40 affected individuals. Stool samples from all affected patients, healthy control individuals (n = 20), and leftover food specimens (n = 2) were collected for laboratory analysis. All samples were processed using standard microbiological methods, conventional PCR, and real-time PCR assay for the identification of causative agents. Pure bacterial cultures underwent antimicrobial sensitivity testing. Data were analyzed to determine the attack rate, construct the epicurve, and perform spot mapping of the outbreak. Atypical enteropatho genic Escherichia coli (aEPEC) was isolated from all 40 affected individuals and one food specimen. No other significant microbial pathogens, including bacteria, viruses, or parasites, could be detected in the specimens by any method. All the aEPEC isolates were sensitive to most antimicrobials, except ampicillin and ceftazidime. Since all the infected patients consumed the same fermented soybean purchased from a single vendor, and aEPEC was isolated from both the patients and the fermented soybean, this suggests a strong correlation between the aEPEC isolates and the outbreak in Keifang, Mizoram, India, which was transmitted through fermented soybean.
The digitalization of food safety management systems (FSMS) represents a crucial strategy for mitigating persistent pathogen contamination and foodborne disease outbreaks. The ISO 22000-based FSMS, incorporating hazard analysis and critical control points (HACCP), relies on periodic verification, retrospective microbiological testing, and manual records, leading to reactive risk management, delayed corrective actions, and limited adaptability to evolving contamination dynamics. Key Industry 4.0 technologies, including digital twins (DT), the Internet of Things (IoT), artificial intelligence (AI), and blockchain, have individually demonstrated the capacity to simulate pathogen contamination dynamics (DT), monitor pathogens in real-time with high sensitivity and predictive accuracy (IoT), enable predictive risk assessment (AI), and reduce traceback from days to seconds (blockchain). However, empirical applications remain limited, with most studies addressing them individually or in pairwise combinations and focusing primarily on supply chain logistics, authenticity, or quality assurance rather than their convergent role in combating foodborne pathogens and supporting HACCP implementation. Following a PRISMA methodology, this review critically examines the potential of DT-centered integration of IoT, AI, and blockchain for pathogen-focused FSMS, which remains underexplored. A unified DT-centered framework linking IoT-based sensing, AI-driven predictive analytics, and blockchain-enabled traceability enables continuous monitoring of critical process and environmental parameters, simulation of contamination dynamics, early risk detection, predictive risk assessment, and enhanced traceability. However, widespread implementation depends on addressing challenges, including heterogeneous data synchronization, interoperability, validation, cybersecurity, implementation costs, and regulatory alignment. Overall, this study provides a pathogen-focused assessment of DT-based systems and outlines future directions for building an intelligent farm-to-fork FSMS.
Religious mass gatherings (MGs) such as Hajj, Umrah, Arba'een and Kumbh Mela attract tens to hundreds of millions of participants annually, posing unique challenges for infectious disease control, surveillance and health system preparedness. We conducted a structured comparative analysis of these four MGs, evaluating attendance scale, participant demographics, governance structures, epidemiological risks and public health response strategies. Data were synthesized from international public health reports, peer-reviewed literature and national MG health policies. Hajj attracts 1.8-3 million pilgrims annually from > 180 countries, while Umrah draws ~ 30 million yearly with similar global diversity. Arba'een hosts 20-22 million participants mainly from Iraq, Iran and neighbouring nations. Kumbh Mela peaks at tens of millions overall, exceeding 50 million on high days, predominantly Indian nationals plus international visitors. Common threats across these mass gatherings include respiratory pathogens, water- and food-borne illnesses and vaccine-preventable diseases, amplified by crowding and diverse origins. Successful public health responses emphasize pre-event vaccination (e.g. meningococcal ACWY for Saudi MGs), real-time surveillance systems and multisectoral coordination among stakeholders. Comparable yet distinct, these MGs require tailored risk-based frameworks emphasizing leadership, surveillance, prevention and surge capacity. Our analysis proposes a unified model to strengthen global MG policies and practices, enhancing health security for future events.
Foodborne Diseases (FBD) are a significant public health issue worldwide, affecting both low and high income countries with over 600 million cases reported in a year. This study provided the first estimate of the economic cost of FBD in Singapore. The incidence of FBD was first estimated using local data sources. We estimated the cases in the community by applying relevant foodborne and underreporting multipliers elicited through expert opinion and the existing literature. The direct and indirect costs associated with the cases were used to estimate the total economic cost due to FBD from known pathogens and gastroenteritis. Direct costs included both inpatient and primary healthcare utilization. Indirect costs included productivity losses due to absenteeism and wage loss due to premature mortality. There were 6080 cases of FBD from known pathogens, with a total societal cost of $15.1 million). There were 184 000 cases of unspecified gastroenteritis, which cost $67.1 million. Nontyphoidal Salmonella contributed the most significant number of cases (n = 2050) and the highest societal cost ($8.12 million) among the known pathogens, whereas Listeria had the highest societal cost per case ($49 900). Our study showed similar results to studies conducted in other countries. This study estimates the true economic burden of FBD. It would help policy makers prioritize FBD so that targeted preventive measures can be implemented to reduce the costs associated with FBD, which pose significant economic costs to society.
Foodborne salmonellosis remains a major global health concern, and the rise of antimicrobial-resistant (AMR) Salmonella strains poses a challenge to both public health and food safety. Whole-genome sequencing (WGS) is a promising alternative to phenotypic antimicrobial susceptibility testing (AST), yet the performance of WGS-based prediction tools requires thorough evaluation across antimicrobial classes. This study assessed the concordance between phenotypic AST and genotypic predictions generated by two WGS pipelines, abritAMR and Staramr. For the analysis, 33 poultry-associated Salmonella enterica isolates were tested against 15 antibiotics spanning seven classes. Antimicrobial resistance determinants were identified in silico and compared with minimum inhibitory concentration-based AST results. Across antibiotics, both pipelines demonstrated strong and comparable performance, achieving similar positive predictive values (abritAMR 83%; Staramr 80%) and negative predictive values (abritAMR 93%; Staramr 95%). High sensitivity and specificity were observed for most antibiotics, including amikacin, ceftriaxone, chloramphenicol, ciprofloxacin, gentamicin, and streptomycin, where both pipelines reached 100% sensitivity with specificity ranging from 88% to 100%. Cohen's κ statistics revealed strong agreement (κ = 0.81-1) for several antibiotics in both abritAMR and Staramr. However, substantial variability was observed for several β-lactams and trimethoprim-sulfamethoxazole, where sensitivity ranged widely (20-100%, depending on the drug and pipeline), reflecting predictive limitations for these classes. Both pipelines met the minimum acceptable performance criteria for 10/15 antibiotics evaluated. Notably, abritAMR demonstrated better overall performance for kanamycin, cefoxitin, and ceftiofur, achieving higher sensitivity and specificity (80-100%) and agreement (κ = 0.81). The disagreements between phenotype and genotype observed may be attributed to factors like gene expression variability, differences in database content, and the complexity of genotype-phenotype relationships. Addressing these gaps may require refinement of pipeline parameters and drug-specific adjustments to improve predictions. Overall, these findings indicate that WGS-based AMR prediction is suitable for several antibiotics, particularly aminoglycosides, fluoroquinolones, and phenicols.
Salmonella species are among the most important foodborne pathogens of public health concern, described by the World Health Organization as high-priority foodborne pathogen causing a wide range of infections. Salmonella harbors numerous virulence genes (VGs) and antimicrobial resistance (AMR) mechanisms that contribute to its pathogenicity and resistance to antimicrobial agents. Therefore, this cross-sectional surveillance study aimed to assess the prevalence, serovar distribution, VGs, and phenotypic AMR profiles of Salmonella isolated from chicken meat collected from broiler slaughterhouses and a processing plant. A total of 859 chicken meat samples (666 from three slaughterhouses and 193 from a processing plant) were collected and analyzed for Salmonella. The pathogen was detected in 188 of 859 samples (21.9%), with multiple serovars identified among the isolates. The prevalence was 10.9% in the processing plant and ranged from 9 to 39% in slaughterhouses. Salmonella Agona (72/188; 38.30%) and Salmonella Molade (18/188; 9.57%) were frequently detected. Among VGs, sitC was the most prevalent, detected in 98.95% of the isolates, while cdtB was the least, found in 12.23% of the isolates. Antimicrobial susceptibility was tested by disk diffusion for most antibiotics and broth microdilution for colistin, following CLSI M100 (2021) guidelines. Most strains were resistant to ampicillin, streptomycin, gentamicin, trimethoprim-sulfamethoxazole, and tetracycline, with resistance rates of 58.4%, 50.0%, 33.5%, 31.9%, and 30.3%, respectively. Colistin showed the lowest resistance rate (7.4%). In contrast, the isolates exhibited high susceptibility to amoxicillin-clavulanic acid (88.8%) and ceftriaxone (86.2%). Overall, multidrug resistance (MDR) was observed in 45.21% of the isolates. MDR was found across several serovars, especially Salmonella Agona. The high prevalence and diversity of Salmonella and its phenotypic AMR in chicken meat indicate widespread contamination in slaughterhouses and processing plants, underscoring the need for stronger control measures and improved regulation of antimicrobial use in poultry production.
Multidrug resistance in foodborne pathogens poses a critical threat to food safety and public health. Enterobacter hormaechei is an emerging pathogen with wide environmental prevalence and is capable of causing severe infections. Bacteriophage-based intervention has gained significant recognition as a sustainable approach to combat foodborne pathogens and address antimicrobial resistance in food production systems. Despite this, research on E. hormaechei-specific phages is facing substantial challenges, primarily due to insufficient phage collections and inadequate genomic characterization of existing isolates. In this work, two novel virulent phages (Ehp-YZU-L3 and Ehp-YZU-L4) were isolated from wastewater samples in Yangzhou. Their morphological, biological, and genomic features were characterized. The two phages belonged to the Myoviridae family, with a latency period (10 and 40 min) and high burst size (192 and 292 plaque-forming unit [PFU]/host cell), and an optimal multiplicity of infection of 0.01. The complete genomic sequences of phages ranged from 163,779 to 170,652 bp and GC content of 39.8 - 40.2%, which consisted of 296 and 275 open reading frames of phage Ehp-YZU-L3 and Ehp-YZU-L4. The absence of both virulence-related genetic elements, antimicrobial resistance, and lysogeny-related genes in two genomes was confirmed. Two phages exhibited strong inhibitory effects against E. hormaechei in pork by a phage-dosage-dependent way, with a reduction range of bacterial counts by 1.73-2.87 Log CFU/g for Ehp-YZU-L3 and 1.96-3.20 log CFU/g for Ehp-YZU-L4 at 37°C for 4 h. These findings demonstrate considerable potential of these two phages for the biocontrol of E. hormaechei contamination in food production systems.
This research examined the in vitro antibacterial, potential antioxidant, and antidiabetic activities of Matricaria chamomilla extract (MCE), Anastatica hierochuntica extract (AHE), and their combination against selected foodborne pathogens. Agar well diffusion, minimum inhibitory concentration (MIC), and minimum bactericidal/fungicidal concentration (MBC/MFC), as well as time-kill effect experiments, were applied to evaluate the antimicrobial characteristics. The combined extract showed better antibacterial qualities than the individual extracts. Results revealed that the tested microbes, including Pseudomonas aeruginosa, Klebsiella pneumoniae, Bacillus cereus, Staphylococcus aureus, and Candida albicans, were among the studied pathogens, with inhibition zones ranging from 24 to 31 mm. By contrast, the inhibition zones of MCE and AHE were 20-26 mm and 19-23 mm, respectively. Furthermore, compared with MCE (150-275 µg/mL; 175-300 µg/mL) and AHE (200-350 µg/mL; 225-375 µg/mL), the combined extract showed reduced MIC (100-200 µg/mL) and MBC/MFC (125-225 µg/mL), indicating even greater antimicrobial efficacy. With an IC50 of 24.44 µg/mL, the combined extract exhibited the greatest radical-scavenging activity in the DPPH experiment, whereas the corresponding values of IC50 for MCE and AHE were 38.19 and 54.49 µg/mL, respectively. The combined extract showed the greatest inhibitory activity against both α-glucosidase and α-amylase. The combined extract showed the strongest and fastest bactericidal action in time-kill tests. Overall, the combined extract demonstrated enhanced, reliable, and repeatable in vitro antibacterial activity, as well as potential antioxidant and antidiabetic properties.
Clostridium perfringens is a widely distributed Gram-positive pathogen associated with a range of foodborne illnesses. In this study, a foodborne outbreak linked to a group meal was investigated, from which 25C. perfringens isolates were recovered from 116 samples collected from food (n = 3), kitchen environmental surfaces (n = 10), food handlers (n = 5), and clinical patients (n = 7). Comprehensive molecular typing using multilocus sequence typing (MLST), core-genome MLST (cgMLST), and single-nucleotide polymorphism (SNP) analysis were performed, and the results showed progressively increasing discriminatory power. MLST divided the C. perfringens isolates into 18 STs, including 14 novel STs. Whole-genome sequencing (WGS) revealed a high level of genetic diversity among the isolates, with 7 enterotoxin-producing type F strains (ST1039) showing notable genetic homogeneity, while 18 type A strains exhibited substantial heterogeneity. High-resolution cgSNP analysis identified a close genomic relationship (15-50 SNP differences) between food isolates-from simmered aged duck soup (hot dish) and salted boiled broad beans (cold dish)-and clinical isolates. Notably, the 15-SNP difference between the duck soup isolate and a clinical isolate suggests a potential epidemiological link to the outbreak cases. This study highlights the superior resolution and accuracy of WGS molecular typing in foodborne outbreak investigations, enabling precise source tracing. It underscores the importance of integrating genomics into routine food safety surveillance and outbreak response to improve public health outcomes.
Brazilian artisanal cheeses are widely valued for their sensory quality and cultural relevance; however, their production frequently involves raw milk and extensive handling, conditions that may favor contamination by Staphylococcus aureus. This retrospective baseline study analyzed artisanal raw-milk cheeses collected between 2014 and 2016 in Southern and Southeastern Brazil and characterized the isolates regarding antimicrobial resistance, biofilm-forming ability, and the presence of virulence and resistance genes. A total of 147 cheese samples were collected from street vendors and markets in five Brazilian states; notably, 93% of the sampling and all S. aureus-positive samples were concentrated in Paraná, São Paulo, and Minas Gerais, Brazil. S. aureus was isolated using selective media and biochemical identification, followed by antimicrobial susceptibility testing according to CLSI guidelines. Molecular analyses were performed to detect enterotoxin genes, the tst gene, the mecA gene, and SCCmec types. Biofilm production was assessed using a microtiter plate assay. Thirteen samples (8.84%) were positive for S. aureus, yielding 25 isolates. Resistance to cefoxitin (FOX) was the most frequent phenotype (20%), followed by resistance to ampicillin, ciprofloxacin, chloramphenicol, and streptomycin (8% each). Two isolates were classified as multidrug-resistant (MDR). Genotypic analysis revealed one mecA-positive isolate (methicillin-resistant S. aureus [MRSA]; SCCmec nontypable), while four FOX-resistant mecA-negative isolates (including three SCCmec type II carriers) were classified as borderline-oxacillin-resistant S. aureus. Biofilm formation was observed among resistant isolates. Among the virulence genes investigated, only one isolate carried the sea gene, and none were positive for tstH. Although the prevalence of mecA-positive MRSA and enterotoxigenic strains was low, the detection of MDR, biofilm-producing S. aureus highlights potential risks to food safety and public health. These findings reinforce the importance of continuous surveillance, molecular monitoring of resistance determinants, and strict hygienic practices in artisanal cheese production.
Fruits and vegetables are vital for nutrition and disease prevention but are prone to contamination throughout the production and postharvest chain. The risk of foodborne infections has increased with the globalization of food supply chains. Foodborne parasites are an emerging public health concern, posing particular risks for populations consuming raw or minimally processed produce. This study investigated parasitic contamination in raw vegetables used to prepare salads sold in Corum Province, Türkiye. From October 2020 to March 2021, 80 locally produced vegetable samples were collected from the central wholesale market. After standardized washing and centrifugation, sediments were examined microscopically, and DNA was extracted for molecular detection of Cryptosporidium, Giardia, Entamoeba, and Blastocystis using polymerase chain reaction (PCR) and sequencing. Overall, 22/80 samples (27.5%) were positive for parasites by both microscopy and PCR. The highest percentages of total samples contaminated were observed in lettuce (8.8%), parsley (6.3%), and arugula (5.0%). Microscopy revealed 13 protozoan cysts and 10 helminth eggs, while PCR identified Blastocystis spp. (15%), Cryptosporidium spp. (3.8%), and Giardia spp. (1.3%). Sequence analysis confirmed Blastocystis ST1, Giardia duodenalis Assemblage A, and Cryptosporidium parvum, all with recognized zoonotic potential. These findings suggest that fresh vegetables in Türkiye may contribute to zoonotic transmission, emphasizing the importance of improved produce hygiene and monitoring. Our findings highlight the urgent need for preventive measures to reduce foodborne parasitic infections in consumers.
Campylobacter is a widespread foodborne pathogen that causes significant contamination across various samples. Rapid and accurate detection of Campylobacter enables timely diagnosis and assessment of contamination, helping to control the spread of the pathogen and ensure food safety. However, the heterogeneity of sample matrices and variability in contamination levels throughout the food supply chain pose substantial challenges to the efficient detection of Campylobacter. In this study, the conventional culture method was improved through the optimization of antibiotic combinations, the A6, A4, and A3 media achieved the highest detection concordance for low (99.04%), medium (98.69%), and high (99.29%) cleanliness samples, respectively. These media also yielded colony recovery counts equivalent to those obtained from antibiotic-free medium, making them suitable for application across the entire chain. The detection sensitivity, specificity, and accuracy of alternative molecular and immunological methods were subsequently evaluated using the optimized culture method for validation. Nucleic acid-based methods generally exhibited high sensitivity (0.98-1.00) and were suitable for use in the upstream and midstream stages, facilitating rapid exclusion of negative samples. Multiplex polymerase chain reaction (PCR) showed good accuracy (0.95-0.98) and allowed identification of contaminating bacteria at the species level, while integration with qPCR facilitated rapid assessment of contamination status of the matrices. Loop-mediated isothermal amplification and colloidal gold immunochromatographic assay offered on-site visualization of results with short detection times and operational simplicity. They also exhibited good accuracy for samples of medium cleanliness (0.80-0.97), making them well-suited to terminal retail applications. Collectively, this study compared and analyzed the applicability of culture-based, nucleic acid-based, and immunological detection methods for samples of varying cleanliness, proposing a comprehensive monitoring strategy to facilitate efficient and accurate detection of Campylobacter in the food supply chain.
Chicken döner is a widely consumed ready-to-eat poultry product that may pose a significant public health risk when prepared under inadequate hygienic conditions. We conducted a retrospective observational study following a large foodborne outbreak linked to chicken döner consumption from a single restaurant in Kocaeli, Türkiye. Epidemiological and clinical data were obtained from hospital records and public health surveillance systems. A total of 1146 emergency department visits were recorded, and 925 unique patients were included after excluding repeat presentations. The median age was 21 years (interquartile range: 16-29), and 63.8% were male. The most common symptoms were nausea (74.5%), vomiting (70.2%), and diarrhea (69.2%). Hospitalization was required in 45 patients (3.9%), with no mortality observed. Hospitalized patients had significantly higher C-reactive protein levels compared with discharged patients (median 76.0 vs. 26.1 mg/L; p = 0.002). Stool cultures, performed in a subset of patients, identified pathogenic organisms in 37.8% of samples, predominantly Salmonella spp. Microbiological analysis of food samples demonstrated multimicrobial contamination, including Salmonella spp., Listeria monocytogenes, Bacillus cereus, and coagulase-positive Staphylococcus. This outbreak represents one of the largest reported chicken döner-associated foodborne events worldwide and highlights the substantial health care burden posed by contaminated ready-to-eat poultry products. Although inflammatory markers such as C-reactive protein were associated with hospitalization, their clinical utility should be interpreted cautiously and in conjunction with overall clinical assessment.
Antimicrobial resistance poses a serious threat to global food safety and poultry production, prompting the need for effective alternatives to conventional antibiotics in food-producing animals. In this study, a recombinant food-grade strain, L. lactis NZ-BB, was engineered to express a fusion antimicrobial peptide (BMAP18-BSN37), and evaluated its probiotic characteristics and antimicrobial efficacy against Salmonella, a major foodborne pathogen in chicken. The recombinant plasmid pUBB was successfully constructed and introduced into L. lactis NZ9000, with optimal peptide expression achieved following Nisin induction (20 ng/mL, 6 h). NZ-BB demonstrated stable plasmid maintenance, high expression levels, and no detectable metabolic burden. In vivo trials using BALB/c murine and 817 strain avian models showed that NZ-BB enhanced body weight gain, supported immune organ development, and improved intestinal barrier integrity through upregulation of tight junction proteins (occludin, claudin-1, ZO-1) and anti-inflammatory cytokines (TGF-β, IL-4), while reducing pro-inflammatory markers (IL-1β, TNF-α, IL-17a). Importantly, oral administration of NZ-BB significantly reduced intestinal and systemic Salmonella burdens, mitigated tissue damage, and restored immune balance in both mice and chicks. Furthermore, NZ-BB regulated the expression of innate immune receptors (e.g., NLRC3) and matrix metalloproteinases (e.g., MMP-1), highlighting its immunomodulatory potential. These results underscore the dual probiotic and antimicrobial functionality of NZ-BB and support its potential use as a food-safe microbial agent to improve animal health and reduce the risk of Salmonella contamination in the food chain.
Contamination of fresh produce with enteric pathogens is a public health concern in low- and middle-income countries (LMICs), especially in informal markets. We assessed the microbiological safety of tomatoes sold at urban markets in Burkina Faso by evaluating the prevalence and concentration of generic Escherichia coli and nontyphoidal Salmonella enterica. Tomatoes were visually categorized into three levels by the amount of damage on tomato surfaces (intact, slightly damaged, severely damaged), and samples were analyzed using a pooled sampling design through Monte Carlo simulations. Mean E. coli prevalence estimates ranged from 4.0 to 18.3%, with both prevalence and mean concentration (3.4-4.1 log10 CFU/tomato) increasing significantly with surface damage. Salmonella enterica prevalence ranged from 0.6 to 2.5%, although prevalence did not vary across damage levels, mean concentrations did increase with tomato damage (1.0-1.5 log10 MPN/tomato). Other market-level factors, including vendor hygiene, ripeness stage, and stall conditions, were not associated with microbial contamination. We did not find a significant correlation between the presence of generic E. coli and Salmonella among market tomatoes. Overall, our findings highlight the utility of visual damage assessment as a proxy for contamination risk. Public health interventions should prioritize damage prevention, improved sorting, vendor training, and consumer education on the safe handling and preparation of tomatoes to reduce foodborne disease burden. Damaged tomatoes were associated with higher levels of microbial contamination and should be cooked and not eaten raw.